FIGURE SUMMARY
Title

New insights into the regulation of cyp3a65 expression in transgenic tg(cyp3a65:GFP) zebrafish embryos

Authors
Erradhouani, C., Piccini, B., Maillot-Marechal, E., Aït-Aïssa, S., Balaguer, P., Coumoul, X., Brion, F.
Source
Full text @ Aquat. Toxicol.
Fig. 1

GFP fluorescence and cyp3a65 mRNA are induced in presence of CLO or TCDD and over-induced when embryos are coexposed to CLO+TCDD. In vivo imaging of tg(cyp3a65:GFP) embryos at 120 hpf. The dashed lines delineate the intestine. Expression in control (CTRL) DMSO 0.02% (A) and exposed embryos to CLO 1.5 µM (B); TCDD 0.05 nM (C) and CLO 1.5 µM + TCDD 0.05 nM (D). (E) corresponds to GFP intensity in individuals (n = 21 embryos minimum per condition) after exposure to CLO 1.5 µM (in blue); TCDD 0.05 nM (in red) and CLO 1.5 µM + TCDD 0.05 nM (in green), control condition (CTRL) is DMSO 0.02% (in black). (F) cyp3a65 mRNA expression in pooled zebrafish embryos (n = 5 embryos/pool) in same condition.

Fig. 2

Clotrimazole-mediated zfPXR activation up-regulates zfahr2 expression and TCDD-mediated zfAhR2 activation up-regulates zfpxr expression. zfpxr mRNA expression in pooled zebrafish embryos (n = 5 embryos/pool) when exposed to (A) CLO and (C) TCDD. zfahr2 mRNA expression in pooled zebrafish embryos (n = 5 embryos/pool) when exposed to (B) CLO and (D) TCDD. Control condition (CTRL) is DMSO 0.01%.

Fig. 3

TCDD promotes strong induction of cyp1a mRNA expression when clotrimazole slightly induces it compared to TCDD. Cyp1a mRNA expression in pooled zebrafish embryos (n = 5 embryos/pool) when exposed to (A) TCDD and (B) CLO.

Fig. 4

GFP intensity in tg(cyp3a65:GFP) embryos exposed to zfPXR and zfAhR2 agonist and antagonist ligands alone or in combination (n = 21 embryos minimum per condition) at 120 hpf after 48 h. Exposure to CLO 1.5 µM; TCDD 0.05 nM; CH22 5 µM and ECN 1 µM alone or in combination. Control condition (CTRL) is DMSO 0.02%.

Fig. 5

Fluticasone propionate promotes fkbp5 expression and inhibits cyp3a65 expression in both GFP intensity (as clobetasol propionate) and mRNA expression. mRNA expression of (A) fkbp5 and (B) cyp3a65 in pooled zebrafish embryos (n = 5 embryos/pool) when exposed to FLU 0.23 µM. GFP intensity in individuals (n = 21 embryos minimum per condition) after exposure to (C) FLU and (D) CLOBE. In (D) FLU is for comparison and TCDD as positive control. Control condition (CTRL) is DMSO 0.01%.

Fig. 6

RU486 reverses inhibited expression of cyp3a65 by fluticasone propionate. (A) In vivo imaging of tg(cyp3a65:GFP) embryos at 120 hpf after 48 h exposure to TCDD 0.05 nM; RU486 1 µM; FLU 0.23 µM and RU486+FLU. TCDD as positive control. Control condition (CTRL) is DMSO 0.02%. The dashed lines delineate the intestine. (B) GFP intensity in individuals (n = 21 embryos minimum per condition) in the same conditions.

Fig. 7

Hypothesis for the regulation of cyp3a65 by zfAhR2, zfPXR and zfGR. While zfAhR2 and zfPXR link to their own response elements in the core of the cyp3a65 promoter, the binding of zfGR to zfHNF4α, itself linked to specific response elements downstream of the cyp3a65 promoter, modulates its expression. zfAhR2 and zfPXR agonist ligands promote the expression of cyp3a65 and zfGR inhibits its constitutive expression. XRE, PXRE and HNF-RE: response elements for zfAhR2, zfPXR and zfHNF4α respectively.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Aquat. Toxicol.
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